Figure 1.
Characterization of HCC827 isogenic cell lines. A, HCC827 isogenic cell lines (HCC827, HCC827-Mock, HCC827-HGF1 and -HGF2, and HCC827 GR5) were cultured overnight in medium containing 10% serum and then incubated for 24 h in serum-free medium, after which the culture supernatants were collected and assayed for HGF with an ELISA. Data are means ± SD from three independent experiments. B, HCC827 isogenic cell lines were cultured in medium containing 10% serum for 72 h in the presence of various concentrations of gefitinib, after which cell viability was assessed as described in Materials and Methods. The number of viable cells is expressed as a percentage of the value for untreated cells. Data are means ± SD from three independent experiments. C, HCC827 isogenic cell lines were incubated for 1 h with or without gefitinib (100 nM) in medium containing 10% serum, after which the cells were lysed and subjected to immunoblot analysis with antibodies to phosphorylated (p-) or total forms of EGFR, MET, AKT, or ERK or with those to β-actin (loading control).